trisodium 3-[(E)-2-(4-{[4-chloro-6-({2-[(4-{[4-(ethenesulfonyl)phenyl]amino}-6-fluoro-1,3,5-triazin-2-yl)amino]propyl}amino)-1,3,5-triazin-2-yl]amino}-5-sulfonaphthalen-1-yl)diazen-1-yl]naphthalene-1,5-disulfonate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

31 July 2006 to 21 September 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40826/A
Batch no.: TZ 5604 BOP 01/06
Expiration date: February 01, 2011
Purity: Content of organic part (Na-salt): approx. 78 %; Oligomers: 13 %; Main component: approx. 48 %
Solubility in water: Approx. >50 g/L at room temperature
Stability in water: Max. 7 days at room temperature
pH: 7.6 (1 g/L)
Aggregate state/physical form at room temperature: Solid (orange powder)
Storage conditions: At room temperature at about 20 °C, away from direct sunlight
Specific instructions: Store in desiccator

Analytical monitoring:

yes

Details on sampling:

For the analysis of the actual test item concentration the following samples were taken just before test start: - duplicate samples from the test medium, - duplicate samples from the control. After 48 and 96 hours: - duplicate samples from the test medium, (stability samples) -duplicate samples from the control. All samples were taken from the approximate center of the aquaria without mixing the test medium, and were deep-frozen (at about -20 °C) immediately after sampling. Based on pre-experiments for investigation of the storage stability (without GLP) the test item is stable in the test water under the storage conditions

Vehicle:

no

Details on test solutions:

At the start of the test, the single test concentration of nominal 100 mg/L was freshly prepared by completely dissolving 500 mg of the test item in 5000 mL of test water using stirring for 20 minutes at room temperature. The test medium was freshly prepared just before introduction of the fish (= start of the test).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

The study was performed with zebra fish (Brachydanio rerio). The test fish were obtained from Zoohaus Schaub, CH-4410 Liestal, Switzerland. In accordance with the test guidelines the fish were held in the laboratories of RCC for more than two weeks without any medication. Prior to test start, they were acclimated for one week to the test water and temperature. During holding and acclimatization until one day before the start of the test the fish were fed with a commercial fish diet (Tetra Min® Hauptfutter, supplied by TETRA-Werke, D-49304 Melle, Germany). During holding and acclimatisation the mortality rate in the test fish batch was lower than 5 % and all fish were healthy. From the acclimated test fish batch, 10 fish were measured at the start of the test: The mean body length of the fish was 3.2 ± 0.15 cm (Mean ± SD), the mean body wet weight was 0.33 ± 0.05 g (Mean ± SD).

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

125 mg/L as CaCO3

Test temperature:

22 °C

pH:

7.5

Dissolved oxygen:

8.4-8.5 mg/L

Nominal and measured concentrations:

Nominal 100 mg/L. The analytically determined test item concentrations in the test media varied in the range from 104 to 108 % of the nominal values at the start and the end of the test period.

Details on test conditions:

One glass aquarium with 5 liters test medium was used for each treatment (the single test concentration and the control). The test vessels were labeled with the RCC study number and all necessary additional information to ensure unmistakable identification. At the start of the test 7 fish were introduced into each aquarium in a random order. The loading rate was 0.46 g fish wet weight per liter test medium. The test medium and the control were slightly aerated during the test period. The fish were not fed during the test. A static test without test medium renewal was chosen since in a pre-experiment (without GLP) the test item concentration in the test medium was constant during the test period of 96 hours. Light conditions were 16-h/8-h light/dark with a 30 min. transition period (light intensity approximately 50-500 Lux).

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 22-73

Details on results:

In the control and in the test medium of 100 mg/L all fish survived until the end of the test and no visible abnormalities were observed at the test fish. Therefore, the 96-hour NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours), and the 96-hour LCO of FAT 40826/A to zebra fish were determined to be at least 100 mg/L. The 96-hour NOEC and the 96-hour LC0 might even be higher than this concentration, but concentrations in excess of 100 mg/L have not been tested, according to the guidelines. The 96-hour LOEC (lowest concentration with toxic effects), the 96-hour LC50 and the 96-hour LC100 were clearly higher than 100 mg/L. These values could not be quantified due to the absence of toxicity of FAT 40826/A at the tested concentration. No remarkable observations were made concerning the appearance of the test medium. It was a clear solution colored by the test item throughout the entire test duration.

Validity criteria fulfilled:

yes

Conclusions:

The 96-h LOEC, LC50 and LC100 of FAT 40826/A were >100 mg/L.

Executive summary:

The acute toxicity of the test item FAT 40826/A to zebra fish (Brachydanio rerio) was determined in a 96-hour static test according to the EU method C.1 (1992), and the OECD Guideline 203. A limit test was performed in accordance with the guidelines to demonstrate that the test item has no toxic effect on the test organisms at the nominal concentration of 100 mg/L. Thus, the only concentration tested was nominal 100 mg/L and a control. The analytically determined concentration of the test item FAT 40826/A in the test medium varied in the range from 104 to 108 % of the nominal value. The test item FAT 40826/A was stable under the conditions of the test during the test period of 96 hours. Therefore, all reported biological results are related to the nominal concentration of the test item. In the control and at the test concentration of 100 mg/L no mortality or other visible abnormalities were determined at the test fish during the test period of 96 hours. Therefore, the 96-hour NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours), and the 96-hour LC0 of FAT 40826/A to zebra fish were determined to be at least 100 mg/L. The NOEC and the LC0 might even be higher than this concentration, but concentrations in excess of 100 mg/L have not been tested, according to the guidelines. The 96-hour LOEC (lowest concentration with toxic effects), the 96-hour LC50 and the 96-hour LC100 were clearly higher than 100 mg/L. These values could not be quantified due to the absence of toxicity of FAT 40826/A at the tested concentration.

Description of key information

The 96-h LC50 of test substance is > 100mg/L in zebra fish.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

In a GLP-compliant study, the acute toxicity of the test item FAT 40826/A to zebra fish (Brachydanio rerio) was determined in a 96-hour static test according to the EU method C.1 (1992), and the OECD Guideline 203. A limit test was performed in accordance with the guidelines to demonstrate that the test item has no toxic effect on the test organisms at the nominal concentration of 100 mg/L. Thus, the only concentration tested was nominal 100 mg/L and a control. The analytically determined concentration of the test item FAT 40826/A in the test medium varied in the range from 104 to 108 % of the nominal value. The test item FAT 40826/A was stable under the conditions of the test during the test period of 96 hours. Therefore, all reported biological results are related to the nominal concentration of the test item. In the control and at the test concentration of 100 mg/L no mortality or other visible abnormalities were determined at the test fish during the test period of 96 hours. Therefore, the 96-hour NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours), and the 96-hour LC0 of FAT 40826/A to zebra fish were determined to be at least 100 mg/L. The NOEC and the LC0 might even be higher than this concentration, but concentrations in excess of 100 mg/L have not been tested, according to the guidelines. The 96-hour LOEC (lowest concentration with toxic effects), the 96-hour LC50 and the 96-hour LC100 were clearly higher than 100 mg/L. These values could not be quantified due to the absence of toxicity of FAT 40826/A at the tested concentration.